Restricted accessResearch articleFirst published online 2013-03
Transplantation of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells or Their Conditioned Medium Prevents Bone Loss in Ovariectomized Nude Mice
Umbilical cord blood (UCB) has recently been recognized as a new source of mesenchymal stem cells (MSCs) for use in stem cell therapy. We studied the effects of systemic injection of human UCB-MSCs and their conditioned medium (CM) on ovariectomy (OVX)-induced bone loss in nude mice. Ten-week-old female nude mice were divided into six groups: Sham-operated mice treated with vehicle (Sham-Vehicle), OVX mice subjected to UCB-MSCs (OVX-MSC), or human dermal fibroblast (OVX-DFB) transplantation, OVX mice treated with UCB-MSC CM (OVX-CM), zoledronate (OVX-Zol), or vehicle (OVX-Vehicle). Although the OVX-Vehicle group exhibited significantly less bone mineral density (BMD) gain compared with the Sham-Vehicle group, transplantation of hUCB-MSCs (OVX-MSC group) has effectively prevented OVX-induced bone mass attenuation. Notably, the OVX-CM group also showed BMD preservation comparable to the OVX-MSC group. In addition, microcomputed tomography analysis demonstrated improved trabecular parameters in both the OVX-MSC and OVX-CM groups compared to the OVX-Vehicle or OVX-DFB group. Histomorphometric analysis showed increased bone formation parameters, accompanied by increased serum procollagen type-I N-telopeptide levels in OVX-MSC and OVX-CM mice. However, cell-trafficking analysis failed to demonstrate engraftment of MSCs in bone tissue 48 h after cell infusion. In vitro, hUCB-MSC CM increased alkaline phosphatase (ALP) activity in human bone marrow-derived MSCs and mRNA expression of collagen type 1, Runx2, osterix, and ALP in C3H10T1/2 cells. Furthermore, hUCB-MSC CM significantly increased survival of osteocyte-like MLO-Y4 cells, while it inhibited osteoclastic differentiation. To summarize, transplantation of hUCB-MSCs could effectively prevent OVX-mediated bone loss in nude mice, which appears to be mediated by a paracrine mechanism rather than direct engraftment of the MSCs.
Get full access to this article
View all access options for this article.
References
1.
BurgeR., Dawson-HughesB., SolomonD.H., WongJ.B., KingA., TostesonA.Incidence and economic burden of osteoporosis-related fractures in the United States, 2005–2025. J Bone Miner Res, 22:465. 2007.
2.
BodyJ.J., BergmannP., BoonenS., BoutsenY., DevogelaerJ.P., GoemaereS.et al.Evidence-based guidelines for the pharmacological treatment of postmenopausal osteoporosis: a consensus document by the Belgian Bone Club. Osteoporos Int, 21:1657. 2010.
3.
SchilcherJ., MichaelssonK., AspenbergP.Bisphosphonate use and atypical fractures of the femoral shaft. N Engl J Med, 364:1728. 2011.
4.
RuggieroS.L.Bisphosphonate-related osteonecrosis of the jaw: an overview. Ann N Y Acad Sci, 1218:38. 2011.
5.
OrwollE.S., ScheeleW.H., PaulS., AdamiS., SyversenU., Diez-PerezA.et al.The effect of teriparatide [human parathyroid hormone (1–34)] therapy on bone density in men with osteoporosis. J Bone Miner Res, 18:9. 2003.
6.
MiyauchiA., MatsumotoT., ShigetaH., TsujimotoM., ThiebaudD., NakamuraT.Effect of teriparatide on bone mineral density and biochemical markers in Japanese women with postmenopausal osteoporosis: a 6-month dose-response study. J Bone Miner Metab, 26:624. 2008.
7.
FlorenzanoF., MinguellJ.J.Autologous mesenchymal stem cell transplantation after acute myocardial infarction. Am J Cardiol, 95:435. 2005.
8.
ImanishiY., SaitoA., KomodaH., Kitagawa-SakakidaS., MiyagawaS., KondohH.et al.Allogenic mesenchymal stem cell transplantation has a therapeutic effect in acute myocardial infarction in rats. J Mol Cell Cardiol, 44:662. 2008.
BhansaliA., UpretiV., KhandelwalN., MarwahaN., GuptaV., SachdevaN.et al.Efficacy of autologous bone marrow-derived stem cell transplantation in patients with type 2 diabetes mellitus. Stem Cells Dev, 18:1407. 2009.
13.
MobasheriA., CsakiC., ClutterbuckA.L., RahmanzadehM., ShakibaeiM.Mesenchymal stem cells in connective tissue engineering and regenerative medicine: applications in cartilage repair and osteoarthritis therapy. Histol Histopathol, 24:347. 2009.
14.
KumarS., WanC., RamaswamyG., ClemensT.L., PonnazhaganS.Mesenchymal stem cells expressing osteogenic and angiogenic factors synergistically enhance bone formation in a mouse model of segmental bone defect. Mol Ther, 18:1026. 2010.
15.
ZhaoM., ZhouJ., LiX., FangT., DaiW., YinW.et al.Repair of bone defect with vascularized tissue engineered bone graft seeded with mesenchymal stem cells in rabbits. Microsurgery, 31:130. 2011.
16.
KimD., ChoS.W., HerS.J., YangJ.Y., KimS.W., KimS.Y.et al.Retrovirus-mediated gene transfer of receptor activator of nuclear factor-kappaB-Fc prevents bone loss in ovariectomized mice. Stem Cells, 24:1798. 2006.
17.
ChoS.W., SunH.J., YangJ.Y., JungJ.Y., AnJ.H., ChoH.Y.et al.Transplantation of mesenchymal stem cells overexpressing RANK-Fc or CXCR4 prevents bone loss in ovariectomized mice. Mol Ther, 17:1979. 2009.
18.
TimmersL., LimS.K., ArslanF., ArmstrongJ.S., HoeferI.E., DoevendansP.A.et al.Reduction of myocardial infarct size by human mesenchymal stem cell conditioned medium. Stem Cell Res, 1:129. 2007.
19.
TimmersL., LimS.K., HoeferI.E., ArslanF., LaiR.C., van OorschotA.A.et al.Human mesenchymal stem cell-conditioned medium improves cardiac function following myocardial infarction. Stem Cell Res, 6:206. 2011.
20.
ChenL., TredgetE.E., WuP.Y., WuY.Paracrine factors of mesenchymal stem cells recruit macrophages and endothelial lineage cells and enhance wound healing. PLoS One, 3:e1886. 2008.
21.
WalterM.N., WrightK.T., FullerH.R., MacNeilS., JohnsonW.E.Mesenchymal stem cell-conditioned medium accelerates skin wound healing: an in vitro study of fibroblast and keratinocyte scratch assays. Exp Cell Res, 316:1271. 2010.
22.
LimH.C., LeeS.T., ChuK., JooK.M., KangL., ImW.S.et al.Neuroprotective effect of neural stem cell-conditioned media in in vitro model of Huntington's disease. Neurosci Lett, 435:175. 2008.
23.
MendesS.C., TibbeJ.M., VeenhofM., BakkerK., BothS., PlatenburgP.P.et al.Bone tissue-engineered implants using human bone marrow stromal cells: effect of culture conditions and donor age. Tissue Eng, 8:911. 2002.
24.
StenderupK., JustesenJ., ClausenC., KassemM.Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. Bone, 33:919. 2003.
25.
StolzingA., JonesE., McGonagleD., ScuttA.Age-related changes in human bone marrow-derived mesenchymal stem cells: consequences for cell therapies. Mech Ageing Dev, 129:163. 2008.
26.
TisatoV., NareshK., GirdlestoneJ., NavarreteC., DazziF.Mesenchymal stem cells of cord blood origin are effective at preventing but not treating graft-versus-host disease. Leukemia, 21:1992. 2007.
27.
MajhailN.S., BrunsteinC.G., TomblynM., ThomasA.J., MillerJ.S., AroraM.et al.Reduced-intensity allogeneic transplant in patients older than 55 years: unrelated umbilical cord blood is safe and effective for patients without a matched related donor. Biol Blood Marrow Transplant, 14:282. 2008.
28.
RebelattoC.K., AguiarA.M., MoretaoM.P., SenegagliaA.C., HansenP., BarchikiF.et al.Dissimilar differentiation of mesenchymal stem cells from bone marrow, umbilical cord blood, and adipose tissue. Exp Biol Med (Maywood), 233:901. 2008.
29.
GuillotP.V., De BariC., Dell'AccioF., KurataH., PolakJ., FiskN.M.Comparative osteogenic transcription profiling of various fetal and adult mesenchymal stem cell sources. Differentiation, 76:946. 2008.
30.
YangS.E., HaC.W., JungM., JinH.J., LeeM., SongH.et al.Mesenchymal stem/progenitor cells developed in cultures from UC blood. Cytotherapy, 6:476. 2004.
31.
LiuY., DingY., MaP., WuZ., DuanH., LiuZ.et al.Enhancement of long-term proliferative capacity of rabbit corneal epithelial cells by embryonic stem cell conditioned medium. Tissue Eng Part C Methods, 16:793. 2010.
32.
YooJ.J., SongW.S., KooK.H., YoonK.S., KimH.J.Osteogenic abilities of bone marrow stromal cells are not defective in patients with osteonecrosis. Int Orthop, 33:867. 2009.
33.
PittengerM.F., MackayA.M., BeckS.C., JaiswalR.K., DouglasR., MoscaJ.D.et al.Multilineage potential of adult human mesenchymal stem cells. Science, 284:143. 1999.
34.
BellinoF.L.Nonprimate animal models of menopause: workshop report. Menopause, 7:14. 2000.
35.
ErbenR.G.Embedding of bone samples in methylmethacrylate: an improved method suitable for bone histomorphometry, histochemistry, and immunohistochemistry. J Histochem Cytochem, 45:307. 1997.
36.
BeckerH.M., ChenM., HayJ.B., CybulskyM.I.Tracking of leukocyte recruitment into tissues of mice by in situ labeling of blood cells with the fluorescent dye CFDA SE. J Immunol Methods, 286:69. 2004.
37.
ChouM.Y., YanD., JafarovT., EverettE.T.Modulation of murine bone marrow-derived CFU-F and CFU-OB by in vivo bisphosphonate and fluoride treatments. Orthodont Craniofac Res, 12:141. 2009.
ChoS.W., YangJ.Y., HerS.J., ChoiH.J., JungJ.Y., SunH.J.et al.Osteoblast-targeted overexpression of PPARgamma inhibited bone mass gain in male mice and accelerated ovariectomy-induced bone loss in female mice. J Bone Miner Res, 26:1939. 2011.
ChoS.W., SunH.J., YangJ.Y., JungJ.Y., ChoiH.J., AnJ.H.et al.Human adipose tissue-derived stromal cell therapy prevents bone loss in ovariectomized nude mouse. Tissue Eng Part A, 18:1067. 2012.
42.
NakanishiC., YamagishiM., YamaharaK., HaginoI., MoriH., SawaY.et al.Activation of cardiac progenitor cells through paracrine effects of mesenchymal stem cells. Biochem Biophys Res Commun, 374:11. 2008.
43.
JungJ., MoonN., AhnJ.Y., OhE.J., KimM., ChoC.S.et al.Mesenchymal stromal cells expanded in human allogenic cord blood serum display higher self-renewal and enhanced osteogenic potential. Stem Cells Dev, 18:559. 2009.
44.
BonewaldL.F.The amazing osteocyte. J Bone Miner Res, 26:229. 2011.
45.
Barragan-AdjemianC., NicolellaD., DusevichV., DallasM.R., EickJ.D., BonewaldL.F.Mechanism by which MLO-A5 late osteoblasts/early osteocytes mineralize in culture: similarities with mineralization of lamellar bone. Calcif Tissue Int, 79:340. 2006.
46.
DunstanC.R., EvansR.A., HillsE., WongS.Y., HiggsR.J.Bone death in hip fracture in the elderly. Calcif Tissue Int, 47:270. 1990.
47.
WongS.Y., EvansR.A., NeedsC., DunstanC.R., HillsE., GarvanJ.The pathogenesis of osteoarthritis of the hip. Evidence for primary osteocyte death. Clin Orthop Relat Res, 214:305. 1987.
48.
WeinsteinR.S., NicholasR.W., ManolagasS.C.Apoptosis of osteocytes in glucocorticoid-induced osteonecrosis of the hip. J Clin Endocrinol Metab, 85:2907. 2000.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
